The first prototype Class III (maximum
containment) biosafety cabinet was
fashioned in 1943 by Hubert Kaempf Jr
April 18, 1955 14 Representatives met at camp
Detrick in Frederick, Maryland.
Purpose: To share knowledge and experiences
regarding bio-safety, chemical, radiological, and
industrial safety issues that were common at the
three principal laboratories of the U.S. Army
1st Biological Safety Conference.
Center for Disease Control (CDC) specifies the
biosafety levels for the U.S.
The standard practices pertain to all levels
Bio-safety Level- Level of the bio-containment
precautions required to isolate dangerous
biological agents in an enclosed facility.
4. BIOHAZARD SYMBOL
Charles Baldwin at
National Cancer Institute
Symbol to be
meaningless” so it could
Blaze orange – most
visible under harsh
6. BSL 1
BIOSAFETY 1 is suitable for work involving
well-characterized agents not known to cause
disease in healthy adult humans, and of minimal
potential hazard to laboratory personnel and the
environment ( CDC 1997).
The laboratory is not necessarily
separated from the general traffic
patterns in the building
7. BSL 1 (CONT)
Work is generally conducted on open bench
tops using standard microbiological
Special containment equipment or facility
designs is not required nor generally used.
Insect and rodent pest control program
should be in effect
Laboratory personnel have specific training
in the procedures conducted in the
laboratory and are supervised by a scientist
with general training in microbiology.
8. BSL 1(CONT)
The following standard special practices,
safety equipment, and facilities apply to agents
assigned to all Biosafety levels.
Standard Microbiological Practices
Access to the laboratory are limited or
restricted when experiments or work with
specimens and cultures are in progress
After handling viable materials and animals
hands must be washed after removing gloves
and before leaving the laboratory.
Eating, drinking, smoking, handling contact
lenses, and applying cosmetics are not
permitted in the work areas where there is
reasonable likelihood of exposure to potentially
infectious materials. Persons who wear contact
lenses in laboratories should also wear goggles
or a face shield. Food is stored outside the work
area in cabinets or refrigerators designated and
used for this purpose only.
Mouth pipetting is prohibited; mechanical
pipetting devices are used.
All procedures are performed carefully to minimize the
creation of splashes or aerosols.
Work surfaces are decontaminated at least once a day and
after any spill of viable material.
All cultures, stocks, and other regulated wastes are
decontaminated before disposal by an approved
Materials to be decontaminated outside of the immediate
laboratory are to be placed in a durable, leak-proof container
and closed for transport from the laboratory.
Materials to be decontaminated at off-site from the laboratory
are packaged in accordance with applicable local, state, and
federal regulations, before removal from the facility.
11. SAFETY EQUIPMENT
Special containment devices or equipment such as a
biological safety cabinet are generally not required for
manipulations of agents assigned to Biosafety Level 1.
It is recommended that laboratory coats, gowns, or uniforms
be worn to prevent contamination or soiling of street clothes.
Gloves should be worn if the skin on the hands is broken or if
a rash exists.
Each laboratory contains a sink for hand
The laboratory is designed so that it can be
easily cleaned. Rugs in laboratories are not
appropriate, and should not be used
because proper decontamination following a
spill extremely difficult to achieve.
Bench tops are impervious to water and
resistant to acids, alkalis, organic solvents,
and moderate heat.
16. BIOSAFETY CONTAINMENT LEVEL 2
Risk Group 2 infectious agents are pathogens
that can cause human or animal disease but,
under normal circumstances, are unlikely to be a
serious hazard to laboratory workers, the
community, livestock, or the environment
Level 2 infections are not considered to be a
serious hazard. They are a moderate individual
risk and limited community risk.
17. BIOSAFETY CONTAINMENT
Laboratory exposures rarely cause infection
leading to serious disease; effective treatment and
preventive measures are available and the risk of
spread is limited.
Examples of infectious agents in this risk level are
E. coli, California encephalitis viruses, many
influenza viruses, some fungi like ringworm;
California encephalitis viruses; human herpes
simplex viruses; many influenza viruses;
Transmissible Gastro-enteritis of swine; Mouse
Hepatitis Virus; and a few parasites.
18. PROCEDURES REQUIRED TO
CONTAIN RISKGROUP 2 ORGANISMS
The laboratory should be separated from all other
A biohazard sign should be present and visible.
All the surfaces in the laboratory should be readily
cleanable and impervious.
An auto clave should be present.
A certified HEPA filtered class 1 or 2 biological cabinet
should be available for the manipulation of organisms.
Gloves and a laboratory coat should be worn at all times
in the laboratory.
20. BIOSAFETY CONTAINMENT
Risk Group 3 infectious agents are pathogens that usually
cause serious human or animal disease, or which can
result in serious economic consequences, but do not
ordinarily spread by casual contact from one individual to
another (high individual risk, low community risk), or that
can be treated by antimicrobial or antiparasitic agents.
Risk Group 3 pathogens include bacteria such as
anthrax, Q Fever, tuberculosis, and viruses such as hanta
viruses, Human immunodeficiency viruses (HIV - all
isolates), eastern and western equine encephalitis viruses.
21. PROCEDURES REQUIRED TO
CONTAIN RISKGROUP 3 ORGANISMS
Laboratory staff must be fully trained in the handling of
pathogenic and other hazardous material, in the use of
safety equipment, disposal techniques, handling of
contaminated waste, and emergency response.
Standard operating procedures must be pasted in visible
Equipment must include an autoclave and a certified
HEPA filtered class II biological safety cabinet
All activities involving infectious materials to be conducted
in biological safety cabinets or other appropriate
combinations of personal protective and physical
22. PROCEDURES REQUIRED TO
CONTAIN RISKGROUP 3
Personal protective equipment should include
head covers and dedicated footwear, gloves,
appropriate respiratory protection
Specialized design and construction of
laboratories, with controlled access double door
entry and body shower.
Air pressure must be negative at all times, no
recirculation, HEPA filtration must be used.
24. BSL- 4
Designed for use with:
Highly toxic/infectious agents
Agents that are at a very high risk for forming infectious aerosols
Life threatening agents
Ex. Ebola, Marburg, Lassa
25. SPECIAL PRACTICES
Only those that work in the BSL-4 lab will be allowed entry
Immuno-compromised individuals such as children and pregnant
women are not allowed to enter
A logbook must be signed each time one comes in or out of the lab
with the date and time
26. SPECIAL PRACTICES
The doors must have a universal hazard label with….
• Description of the infectious agent (s)
• Responsible director
• PPE required such as respirators and any immunization
All lab personnel are trained specially for BSL-4 by lab
27. SPECIAL PRACTICES
Lab personnel must be vaccinated for the agents…
• Present in the lab
• That have potential to be in the lab
Baseline serum samples are collected and stored for serological
• Makes sure the personnel have sufficient antibodies for the pathogens they are
Biosafety manual is adopted for that lab
28. SPECIAL PRACTICES
Entering the lab
• Personnel must remove personal clothing in an outer
room and leave it
• All clothing and PPE is provided and expected to be
Exiting the lab
• Personnel remove lab clothing in the inner room
after taking a decontaminating shower
The lab clothing is autoclaved then cleaned
• They exit through the outer room where their
personal clothes are
29. SPECIAL PRACTICES Supplies/materials are brought into the lab by a
double door autoclave that….
• is secured so that materials can be removed from
the interior door
• is decontaminated after each use
Plastic is substituted for glass when possible
• Eliminate sharps such as capillaries, scalpels,
needles and syringes
• Broken glass should be handled with a dust pan and
• Non-disposable and disposable sharps collected in a
hard container to be transported to an autoclave for
decontamination; disposables are thrown out
30. SPECIAL PRACTICES
Any viable materials transferred from the class III Biological Safety
Cabinet or the lab itself must be put in a primary container and
then in a non- breakable secondary container
• It is then removed by an airlock
• Pressurizes the chamber so that minimal air is emitted when each door opens
one at a time
• Can also decontaminate the containers
31. SPECIAL PRACTICES
Lab equipment is decontaminated after use,
after spills or before any repairs
Any material not related to the project is not
permitted in the laboratory
A system is set up for emergencies such as
exposure to infectious agents and also
employee absenteeism on safety
• Medical care for those with lab acquired illness
• Records are kept for all accidents
32. SAFETY EQUIPMENT
Class III biological safety cabinets for handling
infectious material or….
Class II biological safety cabinet in conjunction
with a positive pressure suit ventilated by a life
BSC must be validated on a 12 month period
The PPE required is a…
• one-piece jumpsuit
• closed toe shoes
• All will be removed in the inner room after showering
33. LABORATORY FACILITY
(SECONDARY BARRIER) BSL-4 labs are housed in separate buildings or isolated
within a building with lower BSL labs
Outer and inner change rooms
Walls, floors and ceilings have sealed internal shells
• keep pests out
• Liquid and heat stabile for decontaminating purposes
Drains have chemical disinfectant traps attached to liquid
waste decontamination system
• All waste must be decontaminated including shower waste
HEPA filters in vents for sterile air
• Filter air exhaust from the class III and II biological safety
• From ventilated positive pressure suits worn in class II safety
34. LABORATORY FACILITY
Windows are break resistant
A dunk tank, fumigation chamber or airlock is
provided for safe passage of materials
outside/inside lab that cannot be
decontaminated in the autoclave
A non-recirculation ventilation system is
provided with directional inward airflow
• Air flows from the outside of the lab to the inside but not vice versa